CN103623240A - Dendrobe processing method and technical screening method of dendrobe - Google Patents
Dendrobe processing method and technical screening method of dendrobe Download PDFInfo
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Abstract
The invention provides a dendrobe processing method and a technical screening method of dendrobe. The dendrobe processing method is characterized by comprising a wine moistening step and a steaming step. The effective components extracted are high in content, the content of dendrobine can reach 0.303%, and the content of polysaccharides can reach 12.3%. The technical screening method is high in precision and accuracy, and interaction among factors can be inspected.
Description
Technical field
The present invention relates to the concocting method of medical material, particularly the concocting method of Herba Dendrobii and processing procedure screening technique.
Technical background
Herba Dendrobii begins to be loaded in < < Sheng Nong's herbal classic > >, classifies as top gradely, has the function of " reinforcing stomach reg fluid, nourishing YIN and clearing away heat ".Contained dry product Herba Dendrobii concocting method of < < Pharmacopoeia of People's Republic of China > > version in 2010 is " remove undesirable root, clean, cutting, is dried ".The prepared slices of Chinese crude drugs concocted specification > > of < < Sichuan Province is identical with pharmacopeia with Herba Dendrobii concocting method in < < Chongqing City prepared slices of Chinese crude drugs concocted specification and standard > >.
Main in Herba Dendrobii containing alkaloid and polysaccharide composition, alkaloid water solublity is bad, is difficult for extracting.Tradition is concocted and is operating as master mainly with personal experience, and the quality of processed product depends on operator's individual subjective judgment, easily causes the unsettled phenomenon of processed product quality.
Summary of the invention
The object of the present invention is to provide a kind of Herba Dendrobii concocting method, the effective ingredient of extraction is as high in the content of dendrobine, dendrobium polysaccharide.
The object of the invention is to realize by following measures:
A Herba Dendrobii concocting method, comprises wine profit and steams step.Wine profit refers to alcoholic solution and infiltrates Herba Dendrobii, steams and refers to steam heating or water proof heating Herba Dendrobii.
In above-mentioned wine profit step, alcoholic solution mass concentration is 45-50%, and the weight ratio of alcoholic solution and Herba Dendrobii is 15-20:100; The time steaming is 6-8 hour.Preferably, above-mentioned ethanol mass concentration is 48%, and the weight ratio of alcoholic solution and Herba Dendrobii is 17:100, and steaming time is 6.78 hours.
Above-mentioned Herba Dendrobii concocting method, comprises the following steps: get clean Herba Dendrobii section and add 45-50% alcoholic solution to carry out wine profit, the weight ratio of ethanol and Herba Dendrobii is 15-20:100, airtight moistening 1-1.5h makes the interior section wettability of medical material even, steam heating steams 6-8 hour, takes out, and dries.
Particularly, above-mentioned Herba Dendrobii concocting method, comprises the following steps:
(1) get stem of Dendrobium and remove undesirable root, clean, cutting is cylindrical or flat columniform section, length 5-10mm, 80 degrees Celsius of dry 2-3h;
(2) wine profit: the Herba Dendrobii that step 1 is obtained is placed in container, add 48% alcoholic solution to mix thoroughly, airtight moistening 1-1.5h, alcoholic solution is penetrated into Herba Dendrobii organization internal slowly, and all by medical material, exhausted as degree, the wettability that makes the Herba Dendrobii section maintenance homogeneous after cutting, reaches inside and outside wettability consistent;
(3) steam: the Herba Dendrobii section after wine profit is packed in rustless steel steamer or porcelain jar, and steam heating 6.78 hours, takes out, and dries.
A kind of another object of the present invention is to provide that precision is high, good stability, reproducible Herba Dendrobii are concocted craft screening method.
A craft screening method for dendrobium wine profit Powder Made by Steamed Method, adopts punctate opacity of the cornea design-effect surface method, it is characterized in that: independent variable is concentration of alcohol, ethanol consumption, steaming time, and efficacy variables is dendrobine content and dendrobium polysaccharide content.
The craft screening method of above-mentioned dendrobium wine profit Powder Made by Steamed Method, comprises the following steps:
Take dendrobine, dendrobium polysaccharide content is index, and each index is all standardized as " the normalizing value " between 0～1, and each index " normalizing value " is asked calculation geometric mean, obtains overall desirability.Formula is: OD=(d1d2 ... dk) 1/k (k is index number), because being, dendrobine and dendrobium polysaccharide content is the bigger the better, so adopt Hassan method to carry out mathematics conversion, ask " normalizing value ", formula is: dmax=(Yi-Ymin)/(Ymax-Ymin).
Use Design-Expert8.0 software to carry out matching to experimental data, equation is OD=5.46-15.9A-16.5B-0.615C+21.1AB+1.01AC+0.452BC+10.9A2+ 27.3B2+0.0226C2 (R2=0.8257p=0.0076).According to response Surface Analysis, draw the surface chart of each dependent variable, see Fig. 2～Fig. 4.
Effect surface analysis learns, along with the increase OD value of steaming time significantly raises, wine concentration and wine consumption affect very not remarkable for OD value, consider definite best processing procedure to be: wine concentration 48%, wine consumption 17%, steaming time 6.78h.Use optimised process to carry out confirmatory experiment three times, dendrobine meansigma methods is 0.303, and predictive value is 0.292.Relative deviation 3.76%.Polysaccharide meansigma methods 12.3, predictive value 11.8.Relative deviation 4.24%.Show that set up model has good predictability.
Beneficial effect
1. Herba Dendrobii concocting method of the present invention, the active constituent content of extraction is high, and dendrobine content can reach 0.303%, and polyoses content can reach 12.3%.
2. Herba Dendrobii concocting method of the present invention, simple to operate, and step, parameter, the easy control of result can quantize, and steady quality be applicable to large-scale industry application, and product is pure, free from admixture, medicine excellent quality.
3. craft screening method of the present invention, investigates for the Relation Parameters in Herba Dendrobii concocting process, has quantized the parameter of several keys, for the foundation of the Standard Operating Procedure of Herba Dendrobii processed product provides foundation.The design of punctate opacity of the cornea that this experiment adopts can well make up that in orthogonal design and uniform Design, test accuracy is inadequate, optimum process condition is inaccurate, reciprocal action such as cannot investigate at the problem between each factor.The optimised process optimizing has quantized influence factor main in concocting process, and the predictive value under optimum process condition and measured value deviation be less.
Accompanying drawing explanation
Reference substance and sample HPLC chromatogram in the 2.1.1 of Fig. 1 embodiment 4; A. reference substance, B. sample.
A and the B effect surface figure to OD value in Fig. 2 embodiment 4
A and the C effect surface figure to OD value in Fig. 3 embodiment 4
B and the C effect surface figure to OD value in Fig. 4 embodiment 4
The specific embodiment
Below by the mode of embodiment, further illustrating the inventive method is specifically described; it is important to point out; following instance is just further detailed the present invention; can not be interpreted as limiting the scope of the invention, the person skilled in the art in this field can do some nonessential improvement and adjustment to the present invention according to foregoing invention content.
Embodiment 1
Herba Dendrobii concocting method, comprises the following steps:
1. get stem of Dendrobium and remove undesirable root, clean, cutting is cylindrical or flat columniform section, length 5mm, 80 degrees Celsius of dry 2h;
2. wine moistens: by the Herba Dendrobii after cutting, put in container, add 50% alcoholic solution to mix thoroughly, the weight ratio of alcoholic solution and Herba Dendrobii is 15:100, airtight moistening 1h, alcoholic solution is penetrated into Herba Dendrobii organization internal slowly, makes the wettability of the Herba Dendrobii section maintenance homogeneous after cutting, and reaches inside and outside humidity consistent.
3. steam: the Herba Dendrobii after wine profit is packed in porcelain jar, and steam heating 6 hours, takes out, and dries.
The active constituent content extracting is high, dendrobine content 0.256%, and polyoses content 11.05%, free from admixture is residual.
Embodiment 2
Herba Dendrobii concocting method, comprises the following steps:
1. get stem of Dendrobium and remove undesirable root, clean, cutting is cylindrical or flat columniform section, length 10mm, 80 degrees Celsius of dry 3h;
2. wine profit: by the Herba Dendrobii after system or cutting only, put in container, add 45% alcoholic solution to mix thoroughly, the weight ratio of alcoholic solution and Herba Dendrobii is 20:100, airtight moistening 1.5h, alcoholic solution is penetrated into Herba Dendrobii organization internal slowly, makes the wettability of the Herba Dendrobii section maintenance homogeneous after cutting, and reaches inside and outside humidity consistent.
3. steam: the Herba Dendrobii after wine profit is packed in rustless steel steamer, and steam heating 8 hours, takes out, and dries.
The active constituent content extracting is high, and dendrobine content can reach 0.278%, and polyoses content can reach 10.18%, and free from admixture is residual.
Embodiment 3
Herba Dendrobii concocting method, comprises the following steps:
1. get stem of Dendrobium and remove undesirable root, clean, cutting is cylindrical or flat columniform section, length 6mm, 80 degrees Celsius of dry 2.5h;
2. wine profit: by the Herba Dendrobii after system or cutting only, put in container, add 48% alcoholic solution to mix thoroughly, the weight ratio of alcoholic solution and Herba Dendrobii is 17:100, airtight vexed 1.2h processed, alcoholic solution is penetrated into Herba Dendrobii organization internal slowly, makes the wettability of the Herba Dendrobii section maintenance homogeneous after cutting, and reaches inside and outside humidity consistent.
3. steam: the Herba Dendrobii after wine profit is packed in rustless steel steamer, and steam heating 6.78 hours, takes out, and dries.
The active constituent content extracting is high, and dendrobine content can reach 0.303%, and polyoses content can reach 12.3%, and free from admixture is residual.
Embodiment 4
1 instrument and reagent
1.1 instrument
Agilent 1200 type high performance liquid chromatograph (G1312B binary pump, G1315C type DAD detector), UV-6000 ultraviolet spectrophotometer (Shanghai Mei Puda Instrument Ltd.), BD61 electronic balance (German SARTORIUS company, sensibility reciprocal is 0.1mg), TGL-18C type centrifuge (Anting Scientific Instrument Factory, Shanghai), RE-52AA type Rotary Evaporators (Shanghai Yarong Biochemical Instrument Plant), HH-4 type digital display thermostat water bath (Jintan City Ke Xi Instrument Ltd.)
1.2 reagent
Reference substance: dendrobine (Man Site bio tech ltd, Chengdu, lot number: MUST-12030205), anhydrous glucose (Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110833-200503)
Medical material: be purchased from the first penetrating judgment of lotus pond medical material market, Chengdu Lu of ，Jing Chengdu University of Traditional Chinese Medicine and award and be accredited as the dry stem of orchid Dendrobium nobile Dendrobium nobile Lindl.
Reagent: high performance liquid chromatography is used chromatographically pure reagent, and all the other are analytical reagent.
2. experimental technique
The assay of 2.1 dendrobines
2.1.1 chromatographic condition
The secondary C18 chromatographic column of chromatographic column Diamonsil diamond (4.6mm * 250mm, 5 μ m), mobile phase acetonitrile-water (0.1% triethylamine) carries out gradient elution (0～30min acetonitrile 30～80%; 30～40min acetonitrile 80%)
[3], flow velocity 1mLmin-1; Detect wavelength 240nm; 30 ℃ of column temperatures; Sample size 10 μ L, HPLC figure is shown in Fig. 1.
2.1.2 the preparation of need testing solution
Get this product powder (crossing sieve No. three) 0.5g, accurately weighed, add after 2mL ammonia infiltrates 30min and add chloroform 50mL, weigh.Reflux, extract, 2h, supplies weightlessness with chloroform.Precision measures subsequent filtrate 10mL, and decompression and solvent recovery, to dry, adds dissolve with methanol, and precision proceeds in 2mL measuring bottle, adds methanol to scale, shake up, and 0.45 μ m filtering with microporous membrane, close plug, standby.
2.1.3 the preparation of standard curve
It is appropriate that precision takes dendrobine reference substance, adds methanol and make every 1ml containing the solution of dendrobine 0.168mg.Draw respectively 2,4,6,8,10,20 μ L, according to 2.2.2 chromatographic condition, carry out HPLC analysis, with peak area and sample size, carry out linear regression, obtain dendrobine linear equation and be respectively: Y=0.0008X-0.213 (R2=0.9998); The range of linearity is 0.336～3.36 μ g.
2.1.4 precision test is got Herba Dendrobii sample, by " 2.1.2 " below legal system available test sample solution, and continuous sample introduction 6 times, the RSD of dendrobine peak area (n=6) is 1.24%.
2.1.5 stability test
Get Herba Dendrobii sample, by " 2.1.2 " below legal system available test sample solution, respectively at 0,2,4,6,8,12h sample introduction, measure, the RSD of dendrobine peak area (n=6) is 1.63%.
2.1.6 replica test
Precision take Herba Dendrobii sample 0.5g6 part by " 2.1.2 " below legal system available test sample solution after sample introduction measure, dendrobine content RSD (n=6) is 1.06%.
2.1.7 average recovery test is accurate takes the Herba Dendrobii sample powder 0.5g that predicts content, and parallel 6 parts, add dendrobine reference substance solution appropriate, by " 2.1.2 " below legal system, for test solution, sample introduction is measured.Calculating dendrobine average recovery rate (n=6) is that 97.62%, RSD is 1.45%.
2.2 dendrobium polysaccharide assays
2.2.1 the preparation of test sample
Get the about 0.5g of Dendrobium (crossing sieve No. three), accurately weighed, add water 200mL, reflux 2h, let cool, be transferred in 250ml measuring bottle, with a small amount of moisture time washing container, washing liquid is incorporated in same measuring bottle, add water to scale, shake up, filter, precision measures subsequent filtrate 10mL, put in centrifuge tube, precision adds dehydrated alcohol 50ml, shake up, cold preservation 1h, take out, centrifugal (rotating speed be per minute 4000 turn) 20 minutes, abandoning supernatant, precipitation adds 80% washing with alcohol 2 times, each 40ml, centrifugal, abandoning supernatant, precipitation heating water dissolves, be transferred in 25ml measuring bottle, let cool, add water to scale, shake up, obtain.
2.2.2 the preparation of standard curve
It is appropriate that precision takes anhydrous glucose reference substance, and adding distil water is made 1mL containing the solution of anhydrous glucose 0.1016mg.Precision measures reference substance solution 0.2,0.4,0.6,0.8,1.0mL, puts respectively in 10mL tool plug test tube, respectively adds water and mends to 1.0mL, precision adds 5% phenol solution 1mL (facing with preparation), shake up, then precision adds sulphuric acid 5mL, shake up, put in boiling water bath and heat 20min, take out, put cooling 5min in ice bath, take corresponding reagent as blank, at the wavelength place of 488nm, measure absorbance, take concentration as vertical coordinate, absorbance is abscissa, drawing standard curve.Y=0.0967X+0.0006 (R2=0.9971) range of linearity 0.02032～0.1016mgmL-1.
2.2.3 precision test
Get Herba Dendrobii need testing solution 1.0mL, press " 2.2.2 " item METHOD FOR CONTINUOUS DETERMINATION sample solution absorbance from " precision adds 5% phenol solution " down, RSD (n=6) is 1.71%.
2.2.4 stability test
Get Herba Dendrobii need testing solution 1.0mL, by " 2.2.2 " item is lower, from " precision adds 5% phenol solution ", every 1h, measure the continuous 6h of absorbance, the RSD of result absorbance (n=6) is 1.55%.
2.3.5 replica test
Precision takes Herba Dendrobii sample 0.5g6 part, and by " 2.2.2 " lower polyoses content of measuring from " precision adds 5% phenol solution ", polyoses content RSD (n=6) is 1.90%.
2.3. punctate opacity of the cornea designs and data analysis
2.3.1 experimental design
On the basis in conjunction with document and preliminary experiment, be chosen at dendrobium wine profit and steam influence factor A Chinese liquor concentration main in process, B Chinese liquor consumption, C steaming time for investigating object design parameter in Table 1, use Design Expert8.0 software to be optimized.Using dendrobine content Y1, polyoses content Y2OD value as investigating index, the results are shown in Table 2.
Table 1 punctate opacity of the cornea design factor water-glass
The design of table 2 punctate opacity of the cornea and result
(noting: 15～No. 20 is parallel laboratory test, with meansigma methods, represents)
2.3.2 date processing
Take dendrobine, dendrobium polysaccharide content is index, and each index is all standardized as " the normalizing value " between 0～1, and each index " normalizing value " is asked calculation geometric mean, obtains overall desirability.Formula is: OD=(d
1d
2d
k)
1/k(k is index number), is the bigger the better because dendrobine and dendrobium polysaccharide content are, so adopt Hassan method to carry out mathematics conversion, asks " normalizing value ", and formula is: d
max=(Y
i-Y
min)/(Y
max-Y
min).
Use Design-Expert8.0 software to carry out matching to experimental data, equation is OD=5.46-15.9A-16.5B-0.615C+21.1AB+1.01AC+0.452BC+10.9A
2+ 27.3B
2+ 0.0226C
2(R
2=0.8257p=0.0076).According to response Surface Analysis, draw the surface chart of each dependent variable, see Fig. 2～Fig. 4.Effect surface analysis learns, along with the increase OD value of steaming time significantly raises, wine concentration and wine consumption affect very not remarkable for OD value, consider definite best processing procedure to be: wine concentration 48%, wine consumption 17%, steaming time 6.78h.Use optimised process to carry out confirmatory experiment three times, dendrobine meansigma methods is 0.303, and predictive value is 0.292.Relative deviation 3.76%.Polysaccharide meansigma methods 12.3, predictive value 11.8.Relative deviation 4.24%.Show that set up model has good predictability.
Claims (7)
1. a Herba Dendrobii concocting method, is characterized in that comprising wine profit and steams step.
2. Herba Dendrobii concocting method as claimed in claim 1, described wine profit is that to adopt mass concentration be that 45-50% alcoholic solution carries out, the weight ratio of alcoholic solution and Herba Dendrobii is 15-20:100, described in time of steaming be 6-8 hour.
3. Herba Dendrobii concocting method as claimed in claim 1 or 2, described ethanol mass concentration is 48%, and the weight ratio of alcoholic solution and Herba Dendrobii is 17:100, and steaming time is 6.78 hours.
4. Herba Dendrobii concocting method as claimed in claim 1, comprise the following steps: get clean Herba Dendrobii and add 45-50% alcoholic solution to carry out wine profit, the weight ratio of ethanol and Herba Dendrobii is 15-20:100, airtight moistening 1-1.5h makes the interior section wettability of medical material even, steam heating steams 6-8 hour, take out, dry.
5. Herba Dendrobii concocting method as claimed in claim 1, comprises the following steps:
(1) get stem of Dendrobium and remove undesirable root, clean, it is 5-10mm that cutting becomes length, 80 degrees Celsius of dry 2-3h;
(2) wine profit: the Herba Dendrobii that step 1 is obtained is placed in container, add 48% alcoholic solution to mix thoroughly, airtight moistening 1-1.5h, alcoholic solution is penetrated into Herba Dendrobii organization internal slowly, the wettability that makes the Herba Dendrobii section maintenance homogeneous after cutting, reaches inside and outside humidity consistent;
(3) steam: the Herba Dendrobii section after wine profit is packed in rustless steel steamer or porcelain jar, and steam heating 6.78 hours, takes out, and dries.
6. the craft screening method of the Herba Dendrobii concocting method as described in as arbitrary in claim 1-5, adopts punctate opacity of the cornea design-effect surface method, it is characterized in that: independent variable is concentration of alcohol, ethanol consumption, steaming time, and efficacy variables is dendrobine content and dendrobium polysaccharide content.
7. the craft screening method of Herba Dendrobii concocting method as claimed in claim 6,, comprise the following steps:
Take dendrobine, dendrobium polysaccharide content is index, and each index is all standardized as " normalizing value " [6] between 0 ~ 1, and each index " normalizing value " is asked calculation geometric mean, obtains overall desirability; Formula is: OD=(d1d2 ... dk) 1/k (k is index number), because being, dendrobine and dendrobium polysaccharide content is the bigger the better, so adopt Hassan method to carry out mathematics conversion, ask " normalizing value ", formula is: dmax=(Yi-Ymin)/and (Ymax-Ymin);
Use Design-Expert 8.0 softwares to carry out matching to experimental data, equation is OD=5.46-15.9A-16.5B-0.615C+21.1AB+1.01AC+0.452BC+10.9A2+ 27.3B2+0.0226C2 (R2=0.8257 p=0.0076); According to response Surface Analysis, draw the surface chart of each dependent variable, analyze effect surface screening technology condition.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104524234A (en) * | 2014-12-29 | 2015-04-22 | 广西玉林容山堂生物科技有限公司 | Dendrobium candidum dry strip and processing method thereof |
| CN106706771A (en) * | 2016-05-21 | 2017-05-24 | 广州今典精方药业有限公司 | Quality standard and preparation process of dendrobium officinale qualitative and quantitative traditional Chinese medicine decoction pieces |
| CN110585317A (en) * | 2019-10-25 | 2019-12-20 | 重庆医药高等专科学校 | Processing method and process screening method of rhizoma pinellinae praeparata |
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| CN102302680A (en) * | 2011-09-16 | 2012-01-04 | 蔡光先 | Processing method capable of reserving all components of dendrobium officinale |
| CN103041186A (en) * | 2013-01-08 | 2013-04-17 | 杭州江南春堂生物科技有限公司 | Novel method for drying fresh dendrobium officinale |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104524234A (en) * | 2014-12-29 | 2015-04-22 | 广西玉林容山堂生物科技有限公司 | Dendrobium candidum dry strip and processing method thereof |
| CN104524234B (en) * | 2014-12-29 | 2018-05-22 | 广西玉林容山堂生物科技有限公司 | A kind of dry item of dendrobium candidum and its concocting method |
| CN106706771A (en) * | 2016-05-21 | 2017-05-24 | 广州今典精方药业有限公司 | Quality standard and preparation process of dendrobium officinale qualitative and quantitative traditional Chinese medicine decoction pieces |
| CN110585317A (en) * | 2019-10-25 | 2019-12-20 | 重庆医药高等专科学校 | Processing method and process screening method of rhizoma pinellinae praeparata |
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Application publication date: 20140312 |